Sample Requirements

If you would like YCGA to prepare samples for sequencing, please see appropriate sample submission requirements below.

If you would like to pool/multiplex/barcode samples to be sequenced in the same flowcell lane, please contact our facility for recommendations on how many samples should be run together. The number of samples to pool will depend on your application and flowcell lane cluster densities. While we strive for a fairly even distribution for each barcode, we cannot guarantee this. When given a high quality sample, we can guarantee a minimum of 170 million clusters per lane. Thank you for understanding.

User-prepared Libraries

If you prepare your own libraries and would like only cluster generation and sequencing services, at least 15ul of undiluted library (>20nM) suspended in EB Buffer should be submitted. Kindly inform us if the DNA was cut with any restriction enzyme or had overhangs before end repair. If you have a modified adaptor or sequencing primer, include the sequencing primer with your submission. We require inhouse quantitation by real-time PCR as an additional service. YCGA has standardized the required concentrations of libraries for optimal cluster generation using YCGA-prepared sequence libraries. We cannot guarantee optimal cluster densities with user-prepared libraries at our standardized picomolar concentration of DNA. Although many user-prepared libraries come close to optimal cluster numbers, some deviate significantly by giving either very low or very high clusters, compromising the number of mappable filtered clusters. In these situations, we still bill for these sub-optimal runs.

Genomic DNA

1 – 5 ug of Purified DNA (5ug recommended) suspended in TE Buffer in a volume not to exceed 50 ul. DNA should be as intact as possible, with an OD260/280 ratio of 1.8-2.

Targeted Exome Capture (Seq-cap)

3 – 5 ug of Purified DNA (5ug recommended) suspended in TE Buffer in a volume not to exceed 50 ul. DNA should be as intact as possible, with an OD260/280 ratio of 1.8-2.

Whole Transcriptome (mRNA-Seq) Profiling

1 - 10 ug of Purified RNA suspended in nuclease-free H2O in a volume less than, or equal to, 50ul. RNA should be as intact as possible, with OD260/280 ratio of 1.8-2 and an OD260/230 ratio of 1.8-2. RNA integrity should be verified via an Agilent Technologies 2100 Bioanalyzer or 1% agarose gel. High-quality RNA will show a 28s rRNA band twice the intensity of the 18S rRNA band.

Small RNA Querying

1 - 10 ug of Purified RNA suspended in nuclease-free H2O in 10 ul volume. RNA should be isolated using a technique that preserves small RNAs (using Qiagen’s miRNeasy kit, for example). RNA should be as intact as possible, with OD260/280 ratio of 1.8-2 and an OD260/230 ratio of 1.8-2. RNA integrity should be verified via an Agilent Technologies 2100 Bioanalyzer or 1% agarose gel. High-quality RNA will show a 28s rRNA band twice the intensity of the 18S rRNA band.